Several materials are known which inactivate serines proteases. Such inactivators can be used for various purposes including affinity labeling of serine proteases to isolate and identify active site sequences, manufacture of plasma proteins and inactivation of certain enzymes in tissue culture. One of the best irreversible inhibitors presently used for serine protease inactivation is diisopropylfluorophosphate (DFP). This material while highly reactive and useful has been found to have drawbacks. It can enter the body of the user by inhalation or cutaneous absorption. Even trace amounts of DFP cause severe miosis while large quantities can cause irreversible inactivation of acetylcholinesterase leading to vomiting, muscle cramps and even death. This makes DFP extremely toxic and dangerous to handle and use. DFP is relatively unstable and can decompose upon exposure to moisture forming hydrogen fluoride which itself causes certain dangers. These toxic properties of DFP have led to its use as a nerve gas in chemical-biological warfare. In contrast the reaction of DFP with serine proteases is often slow and large molar excesses must be used to obtain complete enzyme inactivation.
Phenylmethanesulfonylfluoride, another known serine protease inactivator material is less toxic and more stable than DFP, but like DFP, is also non-selective in that it inhibits both trypsin and chymotrypsin and must be used in large excess with respect to the enzyme to be inactivated. It is known that fast inactivation times and minimized amounts of reactants are preferred in order to speed up processing and to introduce small amounts of materials in certain procedures while minimizing volumes and costs in other procedures.
Certain other sulfonyl fluoride compounds have previously been described for possible use as serine protease inhibitors. Acta Biol. Med. Germ. 28, 577-585 [1972] "Inaktivierung von Trypsin und Thrombin durch 4-Amidinobenzolsulfofluorid und 4-(2-Aminoathyl)-benzosulfofluorid", P. Walsmann, M. Richter and F. Markwardt, describes such materials. However, the materials described are not the specific compounds of this invention and do not have the reactivity and usefulness of the compounds of this invention.